Diagenode

yeast H2BK123ub monoclonal antibody

Antibody ChIP-seq grade icon
Catalog Number
Format
Price
C15200232-10
10 µg
$85.00
  Bulk order
Other format

Monoclonal antibody raised in mouse against the region of yeast histone H2B containing the the ubiquitinylated lysine 123 (H2BK123ub), using a KLH-conjugated synthetic peptide. The antibody also recognizes the human homologue H2BK120ub. 

Lot001
Concentration1 μg/μl
Species reactivityHuman, Yeast
TypeMonoclonal
PurityAffinity purified monoclonal antibody in PBS containing 0.02% azide.
HostMouse
Storage ConditionsStore at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles.
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP * 2 μg/ChIP Fig 1, 2, 3
Western Blotting 1:500 Fig 4, 5

* Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 μg per IP.

  • Validation data

    ChIP

    Figure 1. ChIP results obtained with the Diagenode antibody directed against H2BK123ub
    ChIP assays were performed using the Diagenode antibody against H2BK123ub (Cat. No. C15200232) on sheared chromatin from WT yeast cells, yeast cells with higher steady state levels of H2BK123ub and yeast cells with no ubiquitinylated H2B. Quantitative PCR was performed with primers for the coding region of an active gene. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    ChIP

    Figure 2. ChIP results obtained with the Diagenode monoclonal antibody directed against H2BK123ub
    ChIP assays were performed using human HeLa cells, the Diagenode antibody against H2BK123ub (cat. No. C15200232) and optimized PCR primer sets for qPCR. ChIP was performed with the "iDeal ChIP-seq" kit (cat. No. C01010055) on sheared chromatin from 1,000,000 cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. QPCR was performed with primers for the coding regions of the active CCT5 and EIF2S3 genes, used as positive controls, and for the inactive TSH2B gene and the Sat2 satellite repeat region used as negative controls. Figure 2 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    ChIP-seq

    ChIP-seq

    ChIP-seq

    ChIP-seq

    Figure 3. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H2BK123ub
    ChIP was performed with 2 µg of the Diagenode antibody against H2BK123ub (cat. No. C15200232) on sheared chromatin from 1,000,000 HeLa cells using the “iDeal ChIP-seq” kit as described above. The IP'd DNA was subsequently analysed on an Illumina HiSeq 2000. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 3 shows the peak distribution along the complete sequence and a 3 Mb region of human chromosome 1 (figure 2A and B) and in two genomic regions surrounding the CCT5 and EIF2S3 positive control genes (figure 2C and D).

    WB

    Figure 4. Western blot analysis using the Diagenode antibody directed against H2BK123ub
    Western blot was performed on histone extracts from HeLa cells (15 μg) using the Diagenode antibody against H2BK123ub (Cat. No. C15200232). The antibody was diluted 1:500 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.

    IP

    Figure 5. Western blot analysis using the Diagenode antibody directed against H2BK123ub
    Western blot was performed on whole cell extracts from WT yeast cells (lane 1), yeast cells with higher steady state levels of H2BK123ub (lane 2), none ubiquitinylated H2B (lane 3) and ubiquitinylated H2A (lane 4). The position of the protein of interest is indicated on the right.

  •  Applications
    WB
    Western blot : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies. Learn more about: Load... Read more
    ChIP-qPCR (ab)
    Read more
    IP
    Immunoprecipitation Read more
  •  Documents
    yeast H2BK123ub monoclonal antibody DATASHEET
    Monoclonal antibody raised in mouse against the region of yeast histone H2B containing the the ub...
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    Antibodies you can trust POSTER
    Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar...
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    Epigenetic Antibodies Brochure BROCHURE
    More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e...
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  •  Publications

    How to properly cite this product in your work

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